Cervical cancer, a sexually transmitted disease, is the second leading cause of cancer death in women worldwide. Globally, HPV type 16 is found in over half of all cervical cancers as well as in its precursor lesion, cervical intraepithelial neoplasia (CIN). The HPV E7 gene product is functionally required to maintain the transformed state, and is consistently expressed at high levels in CIN and in cervical cancers, but not in normal tissue. Therefore, it represents a compelling immunotherapeutic target. We have developed a DNA vaccine strategy which targets a detoxified form of HPV16E7 (E7detox) to the MHC class I processing pathway. In the preclinical TC-1 murine model, vaccination with DNA-Sig/E7(detox)/HSP70 elicits a potent E7-specific immune response which is dependent on CD8 T cells but not on CD4 T cells, and furthermore results in significant antitumor effect. Clinical grade vaccine has been procured via the National Cancer Institute Rapid Access to Intervention Development (RAID) program. The overall objective of this proposal is to conduct a phase I/II clinical trial to evaluate this vaccine strategy in terms of safety, toxicity, efficacy, and immune activation in women with CIN2/3. Strengths of this proposal include 1) a multidisciplinary team of investigators with a longstanding commitment and track record together in collaborative research in HPV disease, bringing expertise in tumor immunology, clinical trials, gynecologic pathology, virology, and clinical care of women with CIN, 2) a novel immunotherapeutic intervention with strong biologic rationale and promising preclinical data, and 3) GMP grade drug which has been manufactured by NCI RAID. This project will facilitate translation of a decade of preclinical vaccine development into a potentially effective immunotherapy for HPV-associated cervical disease. Immunological parameters found to be relevant to therapeutic outcomes may be used as predictors of vaccine effects, provide insight into the mechanism of these effects, and inform the future generation of strategies for therapeutic HPV vaccines. [unreadable] [unreadable]